TY - JOUR
T1 - Effect of ischaemic preconditioning on genomic response to cerebral ischaemia
T2 - Similarity to neuroprotective strategies in hibernation and hypoxia-tolerant states
AU - Stenzel-Poore, Mary P.
AU - Stevens, Susan L.
AU - Xiong, Zhigang
AU - Lessov, Nikola S.
AU - Harrington, Christina A.
AU - Mori, Motomi
AU - Meller, Robert
AU - Rosenzweig, Holly L.
AU - Tobar, Eric
AU - Shaw, Tatyana E.
AU - Chu, Xiangping
AU - Simon, Roger P.
N1 - Funding Information:
We thank Yi-Ching Hsieh for work on statistical analysis, Martha Johnson for insightful discussion throughout the project, and Clara Schindler for technical assistance. Microarray assays were done in the Affymetrix Microarray Core of the OHSU Gene Microarray Shared Resource; data analysis was provided through the Bioinformatics and Biostatistics Core. This work was supported by National Institutes of Health grants NS39492 (MS-P), NS35965 (RPS), and NS24728 (RPS).
PY - 2003/9/27
Y1 - 2003/9/27
N2 - Background. Molecular mechanisms of neuroprotection that lead to ischaemic tolerance are incompletely understood. Identification of genes involved in the process would provide insight into cell survival and therapeutic approaches for stroke. We developed a mouse model of neuroprotection in stroke and did gene expression profiling to identify potential neuroprotective genes and their associated pathways. Methods. Eight mice per condition were subjected to occlusion of the middle cerebral artery for 15 min (preconditioning), 60 min (injurious ischaemia), or preconditioning followed 72 h later by injurious ischaemia. RNA was extracted from the cortical regions of the ischaemic and non-ischaemic hemispheres. Three pools per condition were generated, and RNA was hybridised to oligonucleotide microarrays for comparison of ischaemic and non-ischaemic hemispheres. Real-time PCR and western blots were used to validate results. Follow-up experiments were done to address the biological relevance of findings. Findings. Microarray analysis revealed changes in gene expression with little overlap among the conditions of injurious ischaemia, ischaemic preconditioning, or both. Injurious ischaemia induced upregulation of gene expression; 49 (86%) of 57 genes regulated showed increased expression in the ischaemic hemisphere. By contrast, preconditioning followed by injurious ischaemia resulted in pronounced downregulation; 47 (77%) of 61 regulated genes showed lower expression. Preconditioning resulted in transcriptional changes involved in suppression of metabolic pathways and immune responses, reduction of ion-channel activity, and decreased blood coagulation. Interpretation. Preconditioning reprogrammes the response to ischaemic injury. Similar changes reported by others support an evolutionarily conserved endogenous response to decreased blood flow and oxygen limitation such as seen during hibernation.
AB - Background. Molecular mechanisms of neuroprotection that lead to ischaemic tolerance are incompletely understood. Identification of genes involved in the process would provide insight into cell survival and therapeutic approaches for stroke. We developed a mouse model of neuroprotection in stroke and did gene expression profiling to identify potential neuroprotective genes and their associated pathways. Methods. Eight mice per condition were subjected to occlusion of the middle cerebral artery for 15 min (preconditioning), 60 min (injurious ischaemia), or preconditioning followed 72 h later by injurious ischaemia. RNA was extracted from the cortical regions of the ischaemic and non-ischaemic hemispheres. Three pools per condition were generated, and RNA was hybridised to oligonucleotide microarrays for comparison of ischaemic and non-ischaemic hemispheres. Real-time PCR and western blots were used to validate results. Follow-up experiments were done to address the biological relevance of findings. Findings. Microarray analysis revealed changes in gene expression with little overlap among the conditions of injurious ischaemia, ischaemic preconditioning, or both. Injurious ischaemia induced upregulation of gene expression; 49 (86%) of 57 genes regulated showed increased expression in the ischaemic hemisphere. By contrast, preconditioning followed by injurious ischaemia resulted in pronounced downregulation; 47 (77%) of 61 regulated genes showed lower expression. Preconditioning resulted in transcriptional changes involved in suppression of metabolic pathways and immune responses, reduction of ion-channel activity, and decreased blood coagulation. Interpretation. Preconditioning reprogrammes the response to ischaemic injury. Similar changes reported by others support an evolutionarily conserved endogenous response to decreased blood flow and oxygen limitation such as seen during hibernation.
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U2 - 10.1016/S0140-6736(03)14412-1
DO - 10.1016/S0140-6736(03)14412-1
M3 - Article
C2 - 14522533
AN - SCOPUS:0141425718
SN - 0140-6736
VL - 362
SP - 1028
EP - 1037
JO - Lancet
JF - Lancet
IS - 9389
ER -